Abstract

An aerosol biological collector/analyzer, and method of collecting and analyzing an aerosol sample for diagnosis is provided. In particular, the current invention is directed to an airborne aerosol collection and bacterial analysis system and method, capable of collecting an airborne aerosol sample and preparing it for analysis via aerodynamic shock in a single-step.

Description

[0001] 本发明涉及收集和分析经空气传播的细菌或生物的颗粒的系统；更特别地，涉及以下系统，所述系统包括用于经空气传播的细菌或生物的颗粒的检测的气溶胶收集系统，其可以用于制备用于诊断的样品或直接诊断来自于环境或患者(例如，诸如，肺结核)的细菌感染样品。 [0001] The present invention relates to collection and analysis systems by bacteria or biological particles airborne; more particularly, relates to the following, the system includes means for collecting the aerosol particles is detected airborne bacteria or organism of the system, which may be used for sample preparation for diagnostic or diagnosis of bacterial infection in a sample directly from the environment or patient (e.g., such as tuberculosis) is.

背景技术 Background technique

[0002] 肺结核(TB)是接触性传染病，每年导致200万人死亡。 [0002] Tuberculosis (TB) is a contagious disease, resulting in 2 million deaths per year.每年全世界大概930万人发展TB，其中估计大概440万人未被诊断。 About 930 million people worldwide each year develop TB, of which an estimated 440 million people probably have not been diagnosed.改进TB的诊断可能导致每年拯救全世界大约625000调整的生命(annual adjusted livessaved),和从工业化国家中消灭TB。 Improve diagnosis of TB can lead to save the world approximately 625,000 adjusted life (annual adjusted livessaved) per year, and the eradication of TB from industrialized countries.(见，例如，Center ofDisease Controland Prevention, TB elimination:Trendsin tuberculosis2008, (2009)；和WorldHealth Organization, Diagnostics fortuberculosis:global demand and marketpotential (2006),这些的每一个公开内容均在此以引用并入。) (See, for example, Center ofDisease Controland Prevention, TB elimination: Trendsin tuberculosis2008, (2009); and WorldHealth Organization, Diagnostics fortuberculosis: global demand and marketpotential (2006), each of these disclosures are incorporated herein by reference.)

[0003] 然而所有常见的TB诊断设备，除了X射线，均依赖痰样品收集。 [0003] However, all the usual TB diagnostic equipment, in addition to X-rays, rely on sputum sample collection.典型的，每个患者收集三个样品(每个多于3ml)。 Typically, three samples were collected for each patient (for each of more than 3ml).在一些患者人群中，这个步骤特别的困难。 In some patient populations, especially in this difficult step.(见MBConde等人，American Journal ofRespiratoryand Critical Care Medicine, 162:2238_2240,(2000);和0.D.Schoch 等人，American Journal of Respiratory and Critical CareMedicine, 175:80_86，2006，这些的公开内容在此以引用并入)。 (See MBConde et al., American Journal ofRespiratoryand Critical Care Medicine, 162: 2238_2240, (2000); and 0.D.Schoch et al., American Journal of Respiratory and Critical CareMedicine, 175: 80_86,2006, these disclosures incorporated by reference).例如，在儿童的情况下，需要特殊的步骤如痰诱导(sputum induction)和胃抽吸(gastric aspiration);所有这些对于健康护理提供者和患者都是不愉快的和困难的。 For example, in the case of children, it requires special steps such as induced sputum (sputum induction) and gastric aspiration (gastric aspiration); all of which are unpleasant for health care providers and patients and difficult.(World HealthOrganization,Introduction and diagnosis of tuberculosis in children.1nternational Journalof Tuberculosis and Lung Disease, 10(10):10911097，(2006)，其公开内容在此以引用并入。)在痰诱导不成功的情况下，使用光学纤维的支气管镜检查进行已知为支气管肺泡灌洗(BAL)的程序。 (World HealthOrganization, Introduction and diagnosis of tuberculosis in children.1nternational Journalof Tuberculosis and Lung Disease, 10 (10): 10911097, (2006), the disclosure of which is incorporated herein by reference.) In the case of sputum induction unsuccessful, using an optical fiber known as bronchoscopy for bronchoalveolar lavage (BAL) procedure.(NEDunlap 等人，American Journal of Respiratory and CriticalCare Medicine, 161 (4):1376_1395，(2000)，该公开内容在此以引用并入)。 (NEDunlap et al., American Journal of Respiratory and CriticalCare Medicine, 161 (4): 1376_1395, (2000), the disclosure of which is hereby incorporated by reference).该程序需要医学医生，并且如果支气管镜没有适当的消毒，可能导致感染。 The program requires medical doctor, and if not properly disinfected bronchoscopy, may lead to infection.而且，痰样品通常被唾液污染，其降低痰样品的质量(每ml更少的结核分枝杆菌(Mtuberculosis)细胞)导致低的诊断敏感性。 Also, typically a sputum sample contamination with saliva, sputum samples which reduces the quality (fewer per ml Mycobacterium tuberculosis (Mtuberculosis) cells) results in low diagnostic sensitivity.(M.Sakundarno 等人,BMCPulmonary Medicine, 9 (I):16, (2009),该公开内容在此以引用并入。) (M.Sakundarno et al., BMCPulmonary Medicine, 9 (I): 16, (2009), the disclosure of which is hereby incorporated by reference.)

[0004] 每一个TB诊断测试以它的实用性和表现为特征。 [0004] Each TB diagnostic test for its practicality and performance characteristics.(见例如，WorldHealth Organization, Diagnosticsfor tuberculosis:global demandand marketpotential (2006) ；fforId Health Organization, InternationalJournal ofTuberculosis and Lung Disease,10(10): 10911097, (2006)；和WorldHealthOrganization, Global tuberculosis control:Surveillance, planning,financing(2009)，这些的每一个的公开内容均在此以引用并入)。 (See e.g., WorldHealth Organization, Diagnosticsfor tuberculosis: global demandand marketpotential (2006); fforId Health Organization, InternationalJournal ofTuberculosis and Lung Disease, 10 (10): 10911097, (2006); and WorldHealthOrganization, Global tuberculosis control: Surveillance, planning, financing (2009), each of these disclosures are hereby incorporated by reference).实用性指执行测试的时间、训练、资源、易接受性和花费。 Refers to test the practicality of execution time, training, resources, easy to accept and spend.表现指测试的敏感性和特异性。 Performance refers to the sensitivity and specificity of the test.理想的诊断方法是具有高表现的实用的，然而现在的TB诊断固有地具有其上两者的权衡。 Ideal diagnostic method with high performance and practical, but now the TB diagnosis inherently weigh on both.(见上文WHO引文。)这些诊断包括:放射照相法、细菌学培养、药物敏感性测试培养(DST)、痰涂片显微术(SSM)，和核酸扩增测试(NAAT) ο (见上文WHO 引文；和MDPerkins 和J.Cunningham, The Journaloflnfectious Diseases, 196 (Suppll):S15_27，(2007)，这些公开内容在此以引用并入)。 (See WHO quotation above.) These diagnostics include: radiation photography, bacteriological culture, culture drug sensitivity test (DST), sputum smear microscopy (SSM), and nucleic acid amplification test (NAAT) ο (see WHO citations above; and MDPerkins and J.Cunningham, the Journaloflnfectious Diseases, 196 (Suppll): S15_27, (2007), which disclosure is incorporated herein by reference).放射学(胸部X-射线)是最快的诊断方法之一(< I小时)，但受困于非特异性并且需要在实验室操作的昂贵的设备。 Radiology (chest X- ray) is one of the fastest method of diagnosis (<I h), but suffer from non-specific and requires expensive equipment in the laboratory.细菌培养被认为是具有87%的敏感性的黄金标准，并可以检测痰中低至10-100个细胞/ml。 Bacterial culture is considered to have a sensitivity of 87% of the gold standard, and can be detected in sputum up 10-100 cells / ml.使用DST进一步处理细菌培养物允许鉴定MDR-TB的特异性菌株。 DST further processed using bacterial cultures allowed the identification of specific strains of MDR-TB.然而，获得结果需要在资源密集的实验室环境中耗费2-6个星期。 However, the results obtained takes 2-6 weeks in resource-intensive lab environment.(见GEPfyffer等人，Journal of Clinical Microbiology, 35 (9) =2229-2234, (1997)，这个公开内容在此以引用并入)。 (See GEPfyffer et al., Journal of Clinical Microbiology, 35 (9) = 2229-2234, (1997), the disclosure of which is incorporated herein by reference).SSM是最实用且被广泛使用的测试，因为其相对廉价并简单。 SSM is the most practical and widely used test, because it is relatively cheap and simple.痰样品进行结核分枝杆菌的耐酸性染色。 The samples were stained sputum acid-resistant Mycobacterium tuberculosis.显微镜操作员鉴定为阳性反应(ss+)需要多于10000个细胞/ml的痰。 Microscope operator identified as positive (ss +) require more than 10,000 cells / ml sputum.(见上文引用的WH0(2006)公开。)由于积压，这些测试在一些国家可能花费长达6个月并且需要多次造访。 (See above reference WH0 (2006) open.) Because of the backlog, these tests may take up to six months in some countries and require multiple visits.SSM的敏感性低于60% (在高风险组中低至20%)并且尽管在高流行组中特异性高，但在工业化国家中变差。 SSM sensitivity of less than 60% (as low as 20% in the high-risk group) and high specificity despite the high prevalence group, but deteriorated in industrialized countries.(见上文引用的MDPerkins&J.Cunningham)。 (See above reference MDPerkins & J.Cunningham).NAAT对SSM阳性患者(ss+)具有>95%的敏感性(60_70%对于88_)和高特异性。 NAAT has a SSM-positive patients (ss +)> 95% sensitivity (60_70 88_% for) and high specificity.10-1000个细胞/ml的痰包含足以扩增，杂交并产生阳性信号的DNA。 10-1000 sputum cells / ml containing sufficient amplification, hybridization, and DNA to generate a positive signal.(见上文引用的MDPerkins&J.Cunningham)。 (See above reference MDPerkins & J.Cunningham).NAAT和细菌培养有最高的表现并且是仅有的两个可以检测MDR-TB的测试。 NAAT bacterial culture and have the highest performance and the only two can be detected MDR-TB tests.NAAT由于实验室技术人员进行以痰为基础的样品准备遭受低实用性，并因此在资源有限的实验室中高度可变。 NAAT Because the lab technician to sputum-based sample preparation suffer from low availability, and therefore highly variable in resource-limited laboratories.(见上文引用的MDPerkins&J.Cunningham)。 (See above reference MDPerkins & J.Cunningham).如果样品制备方法是成本有效地自动化的，NAAT可能满足发展国家和工业化国家的优先配置。 If the sample preparation method is cost-effective automation, NAAT may be configured to meet the priorities of developing countries and industrialized countries.

[0005] 另一个挑战医学领域的诊断的问题涉及医院内获得性感染(HAI)。 [0005] Another problem diagnosis of the challenges involved in the medical field-acquired infections (HAI) in hospitals.根据疾病控制与预防中心(CDC)，每年在美国医院发生170万HAI (9.3人每1000个患者日(patient-day)或4.5人每100个进入者)，导致每年约99000人死亡。 Centers for Disease Control and Prevention (CDC), each year 1.7 million HAI (9.3 persons per 1000 patient days (patient-day) or 4.5 people per 100 entrants) in the United States according to the hospital, resulting in about 99,000 deaths each year.(见例如RMKlevens等人，Public Health Reports, 122, (2007),这个公开内容在此以引用并入)。 (See for example RMKlevens et al., Public Health Reports, 122, (2007), the disclosure of which is incorporated herein by reference).反之,估计HAI相当于每年超过$350 亿的医疗花费。 On the contrary, it is estimated HAI equivalent to more than $ 35 billion in health care costs each year.(见RD Scott II, Centers for Disease ControlandPrevention，2009，这个公开内容在此以引用并入)。 (See RD Scott II, Centers for Disease ControlandPrevention, 2009, the disclosure of which is incorporated herein by reference).从公共健康的立场和从医疗花费的成本抑制的角度，越来越多地认识到HAI是一个严重的问题。 From a public health standpoint and from a cost point of view to suppress medical costs, more and more aware of the HAI is a serious problem.

[0006] 这些疾病有三种不同的传播途径:接触、飞沫传播，和空气传播。 [0006] These disorders have three different routes of transmission: contacting, droplets, and airborne.飞沫和空气传播需要可以被吸入而导致感染的气溶胶的形成。 Airborne droplets and may need to be caused to form an aerosol inhaled infections.例如，最普遍吸入的感染之一是肺炎。 For example, one of the most common infection is inhalation pneumonia.而且，医院内获得性肺炎(HAP)占所有HAI的11-15%，其导致不成比例的36%的死亡。 In addition, hospital-acquired pneumonia (HAP) accounts for 11-15% of all HAI, which results in a disproportionate 36% of deaths.的确，约79 %的HAP感染是非设备相关的，且多于80 %的HAP感染是细菌性的。 Indeed, about 79% of non-HAP infection associated equipment, and more than 80% of HAP is a bacterial infection.(见DJWeber等人，Infection Control and Hospital Epidemiology, 28 (12):1361_1366, (2007),这个公开内容在此以引用并入)。 (See DJWeber et al., Infection Control and Hospital Epidemiology, 28 (12): 1361_1366, (2007), the disclosure of which is incorporated herein by reference).由此可以估计，将近160，000ΗΑΙ和23，000死亡是由吸入气溶胶细菌(尤其是肺炎链球菌(S.pneumoniae))导致的。 It can be estimated that nearly 160,000ΗΑΙ and 23,000 deaths are caused by inhalation of the aerosol bacteria (particularly Streptococcus pneumoniae (S.pneumoniae)) caused.而且，TB和多药耐药的肺结核(MDR-TB)还面对引起医院内气溶胶传播的另外的问题。 Moreover, TB and multi-drug resistant tuberculosis (MDR-TB) also face additional problems caused by aerosol spread in hospitals.(见SKSharma和A.Mohan, Chest, 130 (I):261-272，(2006)，这个公开内容在此以引用并入。) (See SKSharma and A.Mohan, Chest, 130 (I): 261-272, (2006), the disclosure of which is incorporated herein by reference.)

[0007] 细菌气溶胶收集器/分析器可以诊断直接来自于患者或者在空气中的污染物，具有提供这些传染原的警告并因此避免进一步的感染的潜力。 [0007] Bacterial aerosol collector / analyzer may be diagnosed directly from the patient or contaminants in the air, has the potential to provide warnings of these infectious agents and thus prevent further infection.然而，提供具有足够敏感性的检测系统的关键的挑战是通过向传感器传递高度浓缩的分析物(例如DNA)来收集和快速扩增细菌气溶胶的低浓度。 However, to provide a sufficient critical challenge sensitive detection system is to collect and rapid amplification of the low concentration of bacteria by aerosol delivery of highly concentrated analyte (e.g., DNA) to the sensor.传统的传感器简单地没有满足提供直接诊断来自于气溶胶来源(例如患者的呼气，咳嗽或喷嚏)或来自于环境的可能的感染或污染风险所需要的要求。 Traditional sensor simply does not meet the direct diagnosis from the aerosol source (such as a patient's breath, coughing or sneezing) or from the requirements to the risk of possible infection or contamination of the environment needs.因此，存在提供细菌的和/或生物的收集器/分析器的需求，所述收集器/分析器用于检测可能的直接来自于患者的感染来源和/或来自于空气传播的细菌来源的污染。 Thus, there is a need to provide a bacteria and / or biological collector / analyzer, the collector / analyzer for detecting a possible direct contamination from sources of infection and / or bacterial origin from the airborne.

[0008] 发明概述 [0008] Summary of the Invention

[0009] 本发明涉及一种气溶胶细菌和/或生物收集器/分析器，其用于诊断患者的呼气道感染或空气中的污染。 [0009] The present invention relates to an aerosol of bacterial and / or biological collector / analyzer for diagnosing a patient expiratory air tract infection or contamination.

[0010] 在一个实施方案中，气溶胶收集和分析系统包括: [0010] In one embodiment, the aerosol collection and analysis system comprising:

[0011] •限定流体通道的患者接口，其中第一终端能够接合患者，以至于患者的至少一部分呼气流出物被所述患者接口捕获作为气体样品； [0011] • Patients fluid passage defining an interface, wherein the first terminal is able to engage the patient, so that at least a portion of the patient breath the patient interface effluent is captured as a sample gas;

[0012] •相互连接管，其将接口和撞击器相互连接； [0012] • each connection pipe interconnecting the interface and the striker;

[0013] •空气动力撞击器，其限定在相互连接管和收集容器之间的流体通道，其中空气动力撞击器应用气体样品和悬浮在所述样品中的生物微粒之间的惯性力差异来收集这些生物微粒，所述生物微粒处于其完整的或者裂解的状态； [0013] • aerodynamic striker, which defines a fluid passageway between the tube and are interconnected collection container, wherein the gas sample application aerodynamic striker and the difference between the inertial force of the suspension of biological particles in the sample to collect these biological particles, the biological particle is in its full state or cleavage;

[0014] •收集容器，其与撞击器连接，并容纳用于保存来自于气体样品的生物微粒的液体介质；和 [0014] • collection container is connected with the striker, and receiving a liquid medium for holding biological particles from a gaseous sample; and

[0015] •泵，其用于产生从接口的第一终端到收集容器的流压，以至于气体样品被推动通过由相互连接管和撞击器限定的流体通道而进入收集容器。 [0015] • a pump for generating a flow pressure from the first terminal to the interface of the collecting container, so that the gas sample is pushed through the tube and connected to each other by a striker passage defining a fluid into the collection receptacle.

[0016] 其中在足够的惯性力被施加于生物微粒以裂解微粒的实施方案中，裂解了的生物微粒的内部组分，例如，诸如，DNA被释放。 [0016] wherein a sufficient inertial force is applied to the biological particles to fine particles cleavage embodiment, the lysis component of the internal biological particles, such as for example, the DNA is released.

[0018] 还在另一个实施方案中，相互连接管包括至少一个辅助补偿入口，其被横切于相互连接管的轴布置，提供至大气的出口。 [0018] In yet another embodiment, each connector comprising at least one auxiliary compensation inlet, which is transverse to the axis of each connecting tube is arranged to provide an outlet to the atmosphere.在一个如此的实施方案中，所述至少一个辅助补偿入口被布置为与相互连接管成大于90度的角。 In one such embodiment, the at least one auxiliary inlet is arranged to compensate for the angle of each connecting tube is greater than 90 degrees.还在另一个如此的实施方案中，过滤器被置于辅助补偿入口的出口处。 In yet another such embodiment, the filter is disposed at the outlet of the auxiliary compensation inlet.还在另一个如此的实施方案中，所述过滤器是高效微粒空气(HEPA)过滤器。 In yet another such embodiment, the filter is a high efficiency particulate air (HEPA) filters.

[0019] 还在另一个实施方案中，撞击器包括渐缩喷嘴和平收集表面，渐缩喷嘴限定撞击器的流体通道，平收集表面被置于所述渐缩喷嘴远端并垂直于流动通道。 [0019] In still another embodiment, the nozzle includes a tapered striker peace collecting surface tapering nozzle defines a fluid passage striker, the flat surface of the collection nozzle is placed in the tapered distal end and perpendicular to the flow passage.在如此的一个实施方案中，所述平收集表面具有被布置于其中并与渐缩喷嘴的流体通道对准的入口孔。 In such an embodiment, the flat surface having a collection inlet aperture is disposed therein and aligned with the tapered nozzle fluid passage.在另一个如此的实施方案中，喷嘴的下游样品压力(P1)和喷嘴的上游样品压力(Ptl)遵照不等Sp1A3ci < 0.53。 In another such embodiment, the sample nozzle downstream pressure (P1) and upstream of the sample nozzle pressure (Ptl) varies in accordance with Sp1A3ci <0.53.还在另一个如此的实施方案中，从喷嘴到收集表面的距离(X)和喷嘴的直径(d)具有下述的比例x/d = 1.2。 In yet another such embodiment, the distance from the nozzle to the collection surface (X) and nozzle diameter (d) has a ratio of x / d = 1.2.还在另一个实施方案中，x/d比例被制定为变量。 In yet another embodiment, x / d ratio is formulated as a variable.还在另一个如此的实施方案中，喷嘴的面对样品的表面是镜面抛光的。 In yet another such embodiment, the surface facing the sample nozzle is mirror-polished.

[0020] 还在另一个实施方案中，所述收集容器向第二终端出口逐渐变小。 [0020] In yet another embodiment, the collecting container tapers towards a second outlet terminal.

[0021] 还在另一个实施方案中，在不打开收集容器的任何其他部分的情况下仍可能进入出口。 [0021] In still another embodiment, the opening without any other part of the container may still be collected into the outlet.在一个如此的实施方案中，可能通过注射器进入出口。 In one such embodiment, the outlet may enter through a syringe.[0022] 还在另一个实施方案中，收集容器的第一终端和液体介质表面的距离是大约4mm。 [0022] In yet another embodiment, the liquid medium from the first terminal and the surface of the collection container is about 4mm.

[0023] 还在另一个实施方案中，系统进一步包括传感器，其能够检测来自于裂解的样品的DNA，所述传感器被布置成与收集容器的第二终端流体连接。 [0023] In still another embodiment, the system further comprises a sensor capable of detecting in a sample from the DNA is cleaved, the sensor is arranged to connect the second terminal with a fluid collection container.

[0024] 还在另一个实施方案中，系统进一步包括能够检测完整的细菌的传感器，所述传感器被布置成与收集容器的第二终端流体连接。 [0024] In still another embodiment, the system further comprising intact bacteria capable of detecting a sensor, the sensor is arranged to connect the second terminal with a fluid collection container.

[0025] 还在另一个实施方案中，系统组件间的相互连接部包括具有阀门的快速分离偶联器，以至于一旦分离阀门自动关闭。 [0025] In still another embodiment, the system interconnecting portion between the valve assembly comprises a quick disconnect coupling device, such that the valve automatically closes once separated.

[0026] 还在另一个实施方案中，系统的组件是可加压灭菌的。 [0026] In still another embodiment, the components of the system is a pressurized sterilized.

[0027] 还在另一个实施方案中，系统的组件对普通灭菌化学品是耐受的。 [0027] In still another embodiment, the components of the system are ordinary chemical sterilization tolerated.

[0028] 还在另一个实施方案中，系统的面对样品的表面由塑料材料形成。 [0028] In still another embodiment, the surface of the sample facing the system formed from a plastic material.

[0029] 还在另一个实施方案中，样品通过选自呼气、咳嗽和打喷嚏的方法形成。 [0029] In yet another embodiment, the sample is selected by a breath, coughing and sneezing forming method.

[0030] 还在另一个实施方案中，液体介质是缓冲液。 [0030] In yet another embodiment, the liquid medium is a buffer.

[0031] 在另一个实施方案中，本发明涉及一种针对细菌感染来收集和分析气溶胶样品的方法，其包括: [0031] In another embodiment, the present invention relates to a method for aerosol samples collected and analyzed for bacterial infection, comprising:

[0034] •施加惯性力于样品，以从样品分离其中的生物微粒，或者裂解其中的任何细菌微粒以释放细胞的内部组分；和 [0034] • inertia force is applied to the sample to separate from a sample wherein the biological particles, or any bacterial lysis to release the microparticles wherein the internal components of the cells; and

[0035] •将完整的颗粒或裂解的微粒的内部组分收集于液体介质中以用于分析。 [0035] • the inner component to pass intact or lysed fine particles in the liquid medium is collected for analysis.

[0036] 还在另一个实施方案中，所述方法包括针对患者的震动流出物补偿系统的压力。 [0036] In still another embodiment, the method comprises a pressure compensation system for shaking effluent patient.

[0037] 还在另一个实施方案中，所述方法包括实时分析细菌污染物的细胞的内部组分。 [0037] In yet another embodiment, the method includes internal components of the cell in real time analysis of bacterial contaminants.

[0038] 还在另一个实施方案中，所述方法包括从流体介质中取回内部组分的一批样品用于分析，而不必拆卸系统。 [0038] In yet another embodiment, the method comprises retrieving a sample from a batch fluid medium for analysis of internal components without dismantling the system.

[0039] 附图简述 [0039] BRIEF DESCRIPTION

[0040] 本发明的这些和其他特征和优势，当与所附数据和图联合考虑时，通过参照下列详细描述将被更好地理解，其中: [0040] The present invention These and other features and advantages, when considered jointly with the appended data and, with reference to the following detailed description will be better understood, in which:

[0042] 图1b和Ic提供按照本发明的气溶胶生物收集器/分析器系统的一个组分的实施方案的不意图； [0042] FIGS. 1b and Ic are not intended to provide an embodiment of a component of the aerosol according to the present invention, the biological collector / analyzer system;

[0043] 图1d提供数据图表，显示按照本发明的气溶胶生物收集器/分析器系统的一个实施方案的流速补偿研究； [0043] Figure 1d provides data chart showing the compensation of the flow rate in accordance with one embodiment of the aerosol collector of the present invention bio / analyzer system;

[0044] 图1e和If提供按照本发明的撞击器系统的实施方案的示意图，其中Ie显示撞击器的示意图，且If显示沿图1e的轴“B”的横截面； [0044] Figure 1e and If according to the embodiment provides a schematic striker system of the present invention, wherein a schematic view of the striker display Ie and If in FIG. 1e along display axis "B" of the cross-section;

[0045] 图2a和2b提供下列的方框图:(a)已有的用于空气传播细菌的检测的气溶胶生物传感器；和(b)按照本发明的气溶胶生物收集器/分析器系统的一个实施方案； [0045] The following provides a block diagram of Figures 2a and 2b: (a) for aerosol conventional biosensor detects airborne bacteria; and (b) a biological aerosol collector according to the present invention / analyzer system implementation plan;

[0046] 图3提供按照本发明的气溶胶生物收集器/分析器系统的一个实施方案的示意图； [0046] FIG. 3 provides a schematic view of one embodiment of the aerosol according to the present invention, the biological collector / analyzer system;

[0047] 图4a和4b提供数据图，显示给予一步输入(a)和脉冲输入(b)的传统系统和本发明的系统的理论反应； [0047] Figures 4a and 4b provide data showing the reaction system theory given input step (a) and the pulse input (b) of the present invention and conventional systems;

[0049] 图6提供按照本发明的一个实施方案的用于细菌感染的检测和分析气溶胶样品方法的流程图。 [0049] Figure 6 provides a flowchart according to an embodiment of the present invention for detecting and analyzing the sample aerosol of bacterial infections.

[0050] 发明详述 [0050] DETAILED DESCRIPTION

[0051] 本发明涉及一种气溶胶生物收集器/分析器系统。 [0051] The present invention relates to a biological collector / analyzer system aerosol.特别的，本发明涉及一种经空气传播的气溶胶收集和细菌分析系统，其能够收集经空气传播的气溶胶样品并通过单一步骤的空气动力的冲击将其制备用于分析。 In particular, the present invention relates to an aerosol system for collecting and analyzing the bacteria spread through the air, it is possible to collect aerosol samples propagating through the air and through the impingement of a single step which aerodynamic prepared for analysis.

[0052] 在图1al和la2中图解地描述了本发明的气溶胶收集和分析系统(10)的一种实施方案的示意图。 [0052] In the schematic diagram in FIG. La2 1al and (10) of one of the embodiments described illustrate the aerosol collection and analysis system of the present invention.如所示，患者(12)佩戴面罩或其他患者接口(14),并且患者有规律地呼气，咳嗽，或打喷嚏。 As illustrated, the patient (12) wearing the mask or other patient interface (14), and the patient regularly breath, coughing, or sneezing.由于患者产生气溶胶飞沫(16)，其通常是大约1-12 μ m，它们通过相互连接管(17)被传输至收集器(18)，并同时从样品中被完整地分离(如图la2所示)或通过空气动力撞击器(20)裂解(如图1al所示)。 Since the patient generating an aerosol droplets (16), which is generally about 1-12 μ m, which is transmitted through the interconnecting pipe (17) to a collector (18), while being completely separated (as shown from the sample LA2 shown) or by a pneumatic striker (20) lysis (as shown in Figure 1al).然后暴露的细胞内DNA和或完整的生物颗粒 Then the exposed cellular DNA or intact biological particles and

(22)被收集于合适的介质中，或者直接被传感器(24)检测，产生阳性或者阴性的读数。 (22) is collected in a suitable medium, or directly by a sensor (24) detecting, generating positive or negative readings.

[0053]总的来说，本发明的气溶胶生物收集器/分析器系统包括四个基本的组件，包括:患者/生物传感器接口(14)，其包括能够接合患者并大量地收集所有来自于患者的空气流的装置；用于相互连接患者/传感器接口和撞击器的装置(17)，其包括管过滤器、流量调节器等，能够提供各组件间的相互连接，和平衡来自于患者的自然震动输入流速和恒定的泵流速；撞击器(20)，其包括喷嘴、冲击盘、收集管等，能够分离和/或裂解悬浮在收集的来自于患者的样品中的任何细菌的细胞；和泵(未显示)，其包括所有必须的管、过滤器、流量调节器等，能够提供稳定和足够的流速通过至患者/生物传感器接口的设备。 [0053] In general, a biological aerosol collector according to the present invention / analyzer system includes four basic components, comprising: a patient / biological sensor interface (14), which can engage the patient and comprising a large number of collected from all an air flow of a patient; a means (17) interconnecting the patient / sensor interface and a striker, comprising a filter tube, flow regulators, etc., capable of providing interconnection between the components, and the balance from the patient Natural vibration input flow rate and a constant pump flow rate; striker (20), which includes a nozzle, an impact plate, the collection tubes, capable of separating and / or cracking suspension of any cell of bacteria in a sample collected from a patient; and pump (not shown), which includes all necessary tubes, filters, flow regulator, etc., can provide a stable and sufficient flow through to the patient / device interface biosensor.尽管下文将会描述本发明的生物传感器系统的每一个组件的可选的实施方案，但要理解，这些可选的实施方案仅仅作为实施例提供，本领域普通技术人员要理解对于基础的气溶胶生物收集器/分析器系统的其他的修改方案和可选方案，不离开本发明的范围。 Although the following will be described for each optional component of the biosensor system according to the present embodiment of the invention, it is to be understood that these alternative embodiments are merely provided as embodiments, those of ordinary skill in the art to be understood based aerosol other modifications and alternatives biological collector / analyzer system, without departing from the scope of the invention.

[0054] 关于系统/患者接口(14)，尽管可以使用任何可以提供患者和气溶胶生物收集器/分析器系统之间的相互连接以收集自然呼气样品的装置，例如，诸如，一个简单的管子，但在一个优选的实施方案中，优选设计以适合安全地且全部地覆盖患者的鼻和嘴的接口。 [0054] For the system / patient interface (14), the interconnection between Although any / analyzer system may provide patients and biological aerosol collector to collect natural breath sample apparatus, such as for example, a simple tube but in a preferred embodiment, preferably designed to fit securely and completely cover the nose and mouth of the patient interface.如此的安全的接口是优选的，不但因为其允许患者在测试过程中规律地咳嗽和呼气，而且因为其防止在测试过程中外部的空气污染物进入系统(可以引起检测到环境的细菌)，和同时防止从患者发出的潜在的传染原逸出系统(导致系统损失和敏感度损失，并可以导致其他患者或医护人员感染)。 Thus secure the interface is preferred, not only because it allows the patient to breath and cough regularly during the test, but also because it prevents the air pollutants in the test system into the outside (environment may cause the bacteria were detected), and at the same time preventing the escape of potentially infectious system emanating from the patient (cause system damage and loss of sensitivity, and can lead to other patients or health care workers infected).在一个如此的实施方案中，接口可能包括一个全脸CPAP (Constant Positive Airway Pressure,持续正气道压力)面罩,例如，诸如，来自于Phillips Respironics的ComfortGel l<ullκ。 In one such embodiment, the interface may include a full-face CPAP (Constant Positive Airway Pressure, continuous positive airway pressure) of the mask, e.g., such as from the ComfortGel l Phillips Respironics <ullκ.尽管不必须，优选地，这样的面罩可以是一次性的以减少使用前后的污染的机会。 Although not necessary, preferably, such a mask may be disposable to reduce contamination before and after use of the opportunity.

[0055] 关于相互连接部(17)，应该理解，在本发明中可以使用任何可以提供患者/生物传感器接口(14)和撞击器(20)之间适当的流体连通的装置，例如，诸如，一段简单的管子。 [0055] For interconnecting portion (17), it should be understood that appropriate means may be provided in fluid communication with the patient / biological sensor interface (14) and the striker (20) between any of the present invention can be used in, for example, such as, a simple piece of pipe.然而，在一个实施方案中，在患者/生物传感器接口和撞击器之间提供了能够平衡来自于患者的自然震动输入流速和恒定的泵流速的装置。 However, in one embodiment, between the patient / biosensor interfaces and striker means is provided in the pump flow rate of the patient's natural vibration input flow rate and equilibrium constant can be from.图1b和Ic示意地显示了一个这样的装置的示例性的实施方案。 FIGS. 1b and Ic schematically shows an exemplary embodiment of such a device.如所示，在这个实施方案中，相互连接部(17)通常包括位于相互连接部的一个终端的连接器(26)，其接合接口(14)的开口；和位于相互连接部的第二终端的连接器(28)，其接合撞击器(20)。 As shown, in this embodiment, each connecting portion (17) generally comprises a connector (26) interconnecting a terminal portion, which engages opening interface (14); and a second terminal portion located interconnecting a connector (28) which engages the striker (20).优选地，连接器包括一个透壁塞(through-wall-plug),其包含可加压灭菌的或灭菌的O形环。 Preferably, the connector plug comprising a permeable wall (through-wall-plug), which comprises an O-ring may be autoclaved or sterilized.在这个实施方案中,补偿装置包括一个辅助的或流动补偿开口(30)，顺其长度在其远终端有一个过滤开口(32)用以提供滤过的空气吸入。 In this embodiment, the compensating means comprising a secondary flow or compensation opening (30), along its length to provide a filtered air intake at its distal end there is a filter opening (32).可以提供任何适当的过滤器，例如，诸如，HEPA过滤器。 It may provide any suitable filter, e.g., such as, the HEPA filter.

[0056] 尽管这个连接可以采用任何适当的形式，在一个示例性的实施方案中，辅助的补偿开口包括三通连接，例如Y形连接(Wye connection),其相对于主相互连接部具有一个大于90度的角度。 [0056] Although the connector may take any suitable form, in one exemplary embodiment, the auxiliary compensation opening comprises a three-way connection, such as Y-shaped connector (Wye connection), which are connected to each other with respect to the main portion has a greater than an angle of 90 degrees.提供补偿管有角度的设计，以随着气流通过过滤器离开，使气流转向大于90度，以最小化气溶胶颗粒损失。 Compensation tube angled design, with the air flow to exit through the filter, so that the flow is turned more than 90 degrees, to minimize the loss of aerosol particles.补偿管中的弯曲防止颗粒由于流体的惯性力而跟随气流。 The compensating tube is bent to prevent the particles due to the inertia force of the fluid stream to follow.尽管图1b仅显示了单个补偿开口(30)，如图1c所示，所述开口由一个或多个不同直径的与多个HEPA过滤器连接的侧管组成，以确保足够的流速用于特殊的应用。 Although FIG. 1b only a single compensation opening (30), as shown in Figure 1c, the openings of one or more tubes of different diameter side connected to a plurality of HEPA filters to ensure a sufficient flow rate for a particular Applications.

[0057] 通过这个补偿开口的流动补偿是重要的。 [0057] With this flow compensating compensation opening is important.图1d和下表I显示最大流速(或临界流速)，在给予一个特定的补偿开口直径的情况下可以被补偿。 Table I and Figure 1d shows the maximum flow rate (or critical flow rate), in a case where, given a certain compensation opening diameter can be compensated.所述直径由患者咳嗽过程中的峰流速决定。 The diameter is determined by the peak flow during patient cough.在图1d中概括了计算值，并且下表在穿过管时零压力下降的假设下计算。 Calcd summarized in Figure 1d, and the lower table is calculated on the assumption of zero when the pressure drop through the tube.

[0058] [0058]

[0060] 作为实施例，提供包括五个具有135度角度和5mm直径的侧管的补偿开口可以保持IOOOlpm的流速。 [0060] As an example, providing a five degree angle and 135 having a diameter of 5mm compensating side tube can be kept open in flow IOOOlpm.这个流速代表咳嗽过程中保守的峰流速。 The flow rate represents the conservative cough peak flow process.在咳嗽过程中，已观察到至高3001pm的男性患者的峰流速。 In the course of cough, it has been observed to peak flow supreme 3001pm of male patients.(见例如Gupta JK, Indoor Air，9:517_525 (2009)，这个公开内容在此以引用并入。)在如此的系统下，通过侧管的颗粒损失可以被计算，为16.5%，然而，其他的84.5%的颗粒在Ilpm的气流中被带至撞击器。 (See for example Gupta JK, Indoor Air, 9: 517_525 (2009), the disclosure of which is incorporated herein by reference.) In such a system, the particles may be calculated by the loss of side tube, 16.5%, however, the other 84.5% of the particles are brought to the striker in Ilpm gas stream.这导致带至撞击器的颗粒浓度的1000倍增加并具有最少的损失。 This leads to 1000-fold to a concentration of particles with a striker and with minimal losses increase.

[0061] 因此，这个补偿开口的存在对于可能的流速和装置的操作具有主要影响。 [0061] Thus, the presence of the compensation opening has a major effect on the flow rate and possible operation of the device.随着吸入和呼出，补偿开口将平衡通过HEPA过滤器开口(32)的流。 With the inhalation and exhalation flow compensation opening is balanced by the HEPA filter openings (32).相反，随着患者咳嗽，补偿开口允许系统避免其自身多余的流速。 Conversely, as the patient cough, compensation opening allow the system to avoid their unwanted flow.因此，在撞击器前放置这样的一个补偿开口允许在患者和撞击器之间使用HEPA滤过的空气进行的流体流速的补偿，明显地增加可能的系统流速。 Thus, placement of such a compensation opening allowing fluid flow rate using the compensated HEPA filtered air between the patient and before the impactor strikes, a system may significantly increase the flow rate.而且，具有HEPA过滤器允许避免外部进入和内部外出的污染。 Furthermore, HEPA filter allows to avoid having to enter the external and internal out-contamination.

[0062] 本发明的系统的独特特点之一是其由撞击器系统(20)组成，所述撞击器系统可以收集完整细胞和/或裂解的细胞。 [0062] One of the unique features of the system of the present invention is that by the impactor system (20), with said striker complete system may collect cells and / or lysed cells.也就是说，撞击器系统能够使空气中的细菌细胞经过一个冲击，并且同时地从气体样品中完整分离细胞，或者裂解细胞和收集碎片。 That is, the system enables the impactor bacterial cells through an air impact, and simultaneously separating intact cells from a gaseous sample, or the lysis of cells and debris collection.然而，DNA自身不随经过一次冲击而碎裂。 However, DNA itself with the passage of time does not impact fragmentation.(见例如K.Teshima等人，Shock Waves,4(6):293-297,(1995)，这个公开内容在此以引用并入)。 (See for example K.Teshima et al., Shock Waves, 4 (6): 293-297, (1995), the disclosure of which is incorporated herein by reference).尽管任何可以传递细菌微粒诸如例如完整细胞和/或产生足以裂解细菌细胞的冲击波的适合的空气动力撞击器均可以与本发明的生物传感器一起使用，但在一个实施方案中，如图1e和If所示，撞击器(20)大致上由渐缩喷嘴(34)组成，气溶胶垂直于具有孔(38)的收集表面(36)流过渐缩喷嘴，引向收集贮器(40)。 While any such fine particles can pass, for example, bacteria can be used with the present invention, the biosensor is suitable aerodynamic striker intact cells and / or lysed bacterial cells to generate sufficient shock waves, but in one embodiment, 1e and If in FIG. , the striker (20) substantially by the converging nozzle (34) composed of an aerosol having a vertical bore (38) of the collection surface (36) flows through the converging nozzle, directed collection reservoir (40).

·[0064] 尽管撞击器的每一个组件可以采用一种形式以至于元件的组合能够在气溶胶样品中产生一个适当强力的冲击以裂解由患者产生的任何细菌细胞。 * [0064] Although each striker assembly may take a form such that a combination of elements capable of generating a strong impact in a suitable aerosol sample to lyse the bacterial cells produced by any patient.特别的，应该提供具有喷嘴(34)的撞击器，喷嘴产生惯性颗粒收集所必需的碰撞流。 In particular, the striker should be provided with a nozzle (34), the nozzle flow inertia collision particulate collection necessary.进而，喷嘴应该优选地相对于收集平台的孔(38)放置，以使组件到达尺寸的要求:x/d = 1.2，其中X是从喷嘴出口到收集管入口的距离，d是喷嘴出口的直径。 Furthermore, the nozzle should preferably be (38) disposed relative to the aperture collect the platform, so that assembly to the required size: x / d = 1.2, wherein X is from the nozzle outlet to the distance from the tube inlet to collect, d is the diameter of the nozzle outlet .而且，喷嘴和孔需要完全地对齐并具有至少I%的耐受度。 Further, the nozzle holes need to completely aligned and having at least I% of tolerance.为减少空气流摩擦，喷嘴可以是镜面抛光的。 To reduce the friction of air flow, the nozzle may be mirror-polished.

[0065] 如上所描述的，如图1al和la2图解所示的，在本发明的系统的操作过程中，进入撞击器(20)的气溶胶(16)流动通过渐缩喷嘴(34)，垂直于收集表面(36)，然后通过孔 [0065] As described above, as shown in FIG la2 1al and illustrated, during operation of the system according to the present invention, the aerosol (16) into the striker (20) flows through the converging nozzle (34), vertical a collecting surface (36), then through the holes

(38)，完整的或冲击过的细菌(42)通过孔(38)进入收集液体(44)。 (38), intact bacteria (42) into a collection or impact over the liquid (44) through the aperture (38).收集DNA于缓冲液中，注入传感器系统的裂解后步骤。 The DNA was collected in a buffer, after the step of injecting the cleavage of the sensor system.

[0066] 尽管适用于直接或者用于之后分析收集样品的任何样品收集器(24)均可以被与撞击器(20)整合，但优选的收集器被设计为容纳用以保存样品的缓冲液介质，并且还被设计为在收集前减速样品细菌颗粒。 [0066] Although applicable to any direct or after a sample collector for collecting a sample of analysis (24) may each be the striker (20) is integrated, but preferably is designed to accommodate the collector medium to save the buffer sample and it is also designed as a deceleration bacteria of the sample prior to collection of particles.在一个实施方案中，例如，收集器具有以下特点: In one embodiment, for example, the collector has the following characteristics:

[0067] •位于收集器室的出口，其允许收集室的直接连接，以至于避免暴露于外部环境。 [0067] • the outlet collector chamber, which allows direct connection to the collection chamber, so avoid exposure to the outside environment.通过这样的方法，收集室可以易于与样品提取器相连，以在不需要分离撞击器的情况下轻松提取样品。 By such a method, the collection chamber may be easily connected to the sample extractor to extract the samples easily without the need for impactor separator.这允许在线测量(连续测量)和易于离线测量(或批测量)。 This allows for on-line measurement (continuous measurement) and measuring off easily (or batch measurement).这样的设计使批收集成为可能，但也可以被通向检测器系统的连续泵系统代替。 This design makes it possible to collect batches, but can also be replaced by a detector system leading to continuous pumping system.在一个如此的实施方案中，样品提取器由灭菌注射器组成。 In one such embodiment, the sample is extracted by a syringe sterilized components.

[0068] •收集室，其相对于出口倾斜，以允许从收集器中移出全部的样品。 [0068] • collecting chamber, which is inclined relative to the outlet, to allow all of the samples were removed from the collector.

[0069] •收集器贮器应该有一定容积和尺寸，以至于当填满时，从样品收集介质表面到收集器的孔/入口的距离优选是〜4mm。 [0069] • collector reservoir volume and there should be a certain size, so that when filled, the distance from the medium surface of the sample collected in the collector aperture / inlet preferably ~4mm.模拟表明细菌颗粒将被以少于lOm/s以4mm间隙从管的入口处收集至液体收集表面,显著地减少样品中任何细菌的死亡率。 Simulation shows that the bacterial particles will be of less than / s to 4mm lOm collected liquid to the gap from the entrance surface of the collection tube, significantly reduced mortality of any bacteria in a sample.[0070] 尽管本领域熟知合适的收集介质，例如，诸如，磷酸盐缓冲盐水(PBS)缓冲液，这些收集介质可以与本发明联合使用，但要理解，本发明还允许整合入实时传感器，例如，诸如Fluidigm' s Access Array™系统和其他,其可以鉴定来自于特异于特定细菌(例如结核分枝杆菌(M tuberculosis)细菌)的内部组分的DNA。 [0070] Although well known in the art suitable collection medium, such as for example, phosphate buffered saline (PBS) buffer, the collection medium can be used in combination with the present invention, it is to be understood that the present invention also allows for integration into the real-time sensor, e.g. such Fluidigm 's Access Array ™ system and others, which can be identified in DNA from the inner component specific for a particular bacteria (e.g. Mycobacterium tuberculosis (M tuberculosis) bacteria) is.如此的传感器可以被整合入上文描述的收集模型，以提供细菌或其他呼气道的生物组分的实时感应。 Such sensors may be integrated into the model described above was collected, to provide real-time sensing of bacteria or other biological components in the expiration channel.

[0071] 上文的讨论没有集中在制造本发明的气溶胶生物收集器/分析器系统所使用的材料。 [0071] The above discussion does not concentrate material of the invention is a biological aerosol collector / analyzer system used.要理解，优选地，所述设备应该与外部的环境隔离，并且如此设计以避免可能导致环境细菌的检测的外部的污染，还避免可能导致系统损失和敏感度损失的样品泄漏，样品泄漏还可能导致疾病的扩散。 Be appreciated that, preferably, the device should be isolated from the environment with the outside, and thus result in design to avoid possible external environment detecting bacterial contamination, but also to avoid the leakage of the sample could result in system losses and loss of sensitivity, the sample may also leak lead to the spread of the disease.诸如，使用在组件间的栓塞和槽上均具有阀门的快速分离偶联器，例如，诸如，Acetal* Quick-Disconnect Coupling Plug( Acetal* 快速分离偶联栓塞)可以允许撞击器的下室在使用前和使用后被分离。 Such as, using a homogenizer with a valve in a rapid separation of the plug and the coupling groove between components, e.g., such as, Acetal * Quick-Disconnect Coupling Plug (Acetal * Rapid Resolution coupling plug) may allow the impactor chamber, in use, after separation and prior to use.如此的布置可以减少外部进入和内部外出的污染，并且优选于帮助在使用前和使用后控制污染。 Such arrangement may reduce external incoming and outgoing internal contamination, and preferably on helping before and after use in pollution control.

[0072] 此外，设备应该是无污染的，并因此应该被设计为经得起使用于其他医学设备的标准灭菌仪器。 [0072] In addition, the device should be non-polluting, and should therefore be designed to withstand use in other medical devices standard sterilization equipment.因此，所有选择的材料应该是可加压灭菌的或应该是购入灭菌的。 Thus, all of the selected materials should be autoclaved or sterilized should be purchased.而且，为最小化样品损失，应该评价使用材料的静电性质，以避免收集带电荷的生物气溶胶。 Furthermore, to minimize sample loss, it should be evaluated using the electrostatic properties of the material, in order to avoid collecting bioaerosols charged.特别是，因为细菌通常是带负电荷的颗粒，不保持静电电荷的塑料对于可能与样品相互作用的部件可能是优选的材料。 In particular, because the bacteria are generally negatively charged particles, electrostatic charge is not maintained for the plastic member may interact with the sample material it may be preferred.

[0073] 尽管上文讨论的没有集中在用于分析气溶胶样品的气溶胶生物收集器/分析器系统，本发明还涉及分析气溶胶样品的方法。 [0073] Although not concentrated in the aerosol collector for the analysis of biological samples of aerosol / analyzer system discussed above, the present invention also relates to a method for analyzing an aerosol sample.如图6中的流程图所示，所述方法包括大概以下步骤: As shown in the flowchart, the method comprising the steps of roughly:

[0074] •收集来自于患者的呼气流出物，包括任何咳嗽或打喷嚏，作为样品； [0074] • exhalation collected in the effluent from a patient, including any coughing or sneezing, as a sample;

[0075] •传输样品至撞击器； [0075] • transfer the sample to the striker;

[0076] •对样品应用惯性力以分离样品中的任何生物微粒和/或裂解其中包含的任何细菌细胞，由此释放其中的任何内部组分；和 [0076] • an inertial force applied to the sample to separate any biological particles in the sample and / or any bacterial cell lysis contained therein, thereby releasing any internal components thereof; and

[0077] •收集完整的微粒和/或内部组分于液体介质中，用于之后的分析。 In a liquid medium, for later analysis [0077] • Complete collected particulates and / or internal components.

[0078] 尽管这些是过程中的基本步骤，应该要理解，可能包括其他步骤以适合上文讨论所描述的设备的操作。 [0078] While these are the basic steps in the process, it should be understood, it may include other steps for operation of the device described hereinabove discussed.例如，在一个实施方案中，所述方法可能还包括补偿来自于患者呼气的流入和流出和来自于由咳嗽和/或打喷嚏导致的任何过度压力的自然震动的系统压力。 For example, in one embodiment, the method may further include compensating exhalation from the patient and from the inflow and outflow of cough and / or any natural vibrations of the system pressure of the excess pressure caused by sneezing.

[0079] 在分析其自身方面，可能通过批方法从液体介质中取出个体样品来分析。 [0079], individual samples may be analyzed removed from the liquid medium in the analysis of its own by the batch method.可选的，可以通过在线分析器和液体介质的相互连接实时完成分析。 Alternatively, the analysis can be accomplished by connecting line analyzer and the liquid medium in real time.

具体实施方式 detailed description

[0080] 在此章节中，提供如何操作气溶胶生物收集器/分析器系统或如何应用所述收集器/分析器系统的若干实施例。 [0080] In this section, how to provide a biological aerosol collector / analyzer system or how to apply several embodiments of the collector / analyzer system.此外，提供按照本发明制造的气溶胶生物收集器/分析器系统的表现与传统系统的表现的对比。 Furthermore, according to the comparative performance of the aerosol performance of the present invention for producing bio-collector is / analyzer system of the conventional system.本领域技术人员将认识到，根据本发明的额外的实施方案涵盖在前述的普通公开的范围内，并且前述的非限制性的实施例没有预期以任何方式的放弃。 Those skilled in the art will recognize that additional embodiments according to the present invention are encompassed within the general scope of the disclosure, and the aforementioned non-limiting embodiment does not in any way contemplated abandoned.

[0081] 实施例1:与传统系统的表现的对比-环境的取样 [0081] Example 1: Comparison of the performance of the conventional system - sampling environment

[0082] 转向本发明的操作与传统气溶胶收集和分析系统的操作的对比，首先应该要理解，接近细胞的内部组分(IC)，主要是DNA，对大多数检测方法是必须的。 Comparison of the operations [0082] of the present invention, the steering operation of the conventional aerosol collection and analysis system, firstly it should be understood that access to the interior components of the cell (IC), mainly a DNA, most detection methods is necessary.(见例如N.Bao和C.Lu., Principles of Bacterial Detection:Biosensors, Recognition Receptorsand Microsystems,pages817-831.Springer, (2008),这个公开内容在此以引用并入)。 (See e.g. N.Bao and C.Lu., Principles of Bacterial Detection: Biosensors, Recognition Receptorsand Microsystems, pages817-831.Springer, (2008), the disclosure of which is incorporated herein by reference).在传统的生物传感器系统中，如图2a的方框图所示，细菌的气溶胶是首先被收集于第一部件 In a conventional biosensor system, the block diagram shown in Figure 2a, the aerosol bacteria are first collected in a first member

(46),然后通过机械系统、加热系统、化学系统、电系统或者激光系统在弟_■部件(48)中裂解。 (46), and then by mechanical systems, heating systems, chemical systems, electrical systems or a laser system cleaved Di _ ■ member (48).然后，裂解产物在下一个部件(未显示)被纯化，以传递靶分子至合适的传感器。 Then, the next lysates member (not shown) is purified to a suitable sensor is transmitted to the target molecule.(对于图2a和2b，要注意，阴影区域表示用于传输分析物至传感器的液体介质，椭圆形和螺旋形分别代表完整的细菌和释放的DNA。还应该注意，在本发明的系统里，DNA释放发生在空气中。) (For Figures 2a and 2b, there is noted that the shaded area represents the liquid medium for the transmission of the analyte to the sensor, elliptical and spiral represent intact bacteria and release DNA. It should also be noted that, in the system of the present invention, DNA release occurs in the air.)

[0083] 简言之，已有的系统(图2a)要求两个分离的并不相连的部件:一个收集器(例如一个旋风分离器(cyclone))，和一个提取器(例如一个声波仪)。 [0083] In brief, the existing system (FIG. 2a) does not require two separate components joined: a collector (e.g. a cyclone (Cyclone)), and an extractor (e.g., a sonicator) .相反，本发明的气溶胶生物收集器/分析器系统(示于图2b的方框图)通过在一个部件(50)中而非多步骤过程在空气中破裂细胞并同时收集它们的IC或DNA而直接传递裂解的产物至已有系统的纯化部件(未显示)。 In contrast, the biological aerosol collector according to the invention / analyzer system (block diagram in Figure 2b) by a multi-step process, rather than ruptured cells in the air at a member (50) and simultaneously collect their DNA directly or IC cleavage product was transferred to an existing purification system components (not shown).换种说法，两个过程(收集&裂解)通过新的收集/撞击器系统而发生在单一步骤中。 Stated differently, the two processes (collect & cleavage) takes place in a single step through a new collector / impactor system.

[0084] 两者的目的是通过细胞内DNA(yi)快速并浓缩的释放而检测经空气传播的肺炎链球菌的浓度变化(Xgl)。 [0084] the purpose is to detect changes in the concentration of Streptococcus pneumoniae propagated through the air (Xgl) by intracellular DNA (yi) and concentrated quick release.在图2a中显示的方法所要求的过程的动力学以下列三个不同的等式(ODE)描述: In the following three different equations (ODE) is described in FIG. 2a dynamics required in the process:

[0088] 其中G和L分别是气体和液体取样流速；下标g和I分别指气相和液相；x是细菌的计数浓度是注入纯化步骤(图2未显示)的DNA的计数浓度；τ是每个过程的停留时间(死容积除以L)和η是气体取样器的收集效率。 [0088] wherein G and L are the gas and liquid sampling flow rate; and g subscripts refer to gas and liquid phases I; x is the concentration of bacteria was counted injection purification step (not shown in FIG. 2) counts concentration of DNA; [tau] residence time of each process (dead volume divided by L), and η is the collection efficiency of the gas sampler.

[0089] 在这个计算中进行了很多假设。 [0089] Many assumptions made in this calculation.例如，为这个计算假设每个细胞存在一个DNA分子。 For example, this calculation is assumed that there is a DNA molecule per cell.相对于空气中的将导致感染的细菌的临界浓度(即Xgl > I导致感染)，标准化所有浓度(无量纲)。 Relative to the air will cause the critical concentration of bacterial infections (i.e. Xgl> I lead to infection), all normalized concentration (dimensionless).假设细胞的破裂在一个连续搅拌罐反应器中(CSTR)是第一个顺序反应，具有反应常熟k。 Suppose ruptured cells in a continuous stirred tank reactor (CSTR) is a first order reaction, the reaction Changshu having k.(见例如JAAsenj0.Separation Processes in Biotechnology.MarcelDekker, New York, lstedition, (1990),这个公开内容在此以引用并入。) (See e.g. JAAsenj0.Separation Processes in Biotechnology.MarcelDekker, New York, lstedition, (1990), the disclosure of which is incorporated herein by reference.)

[0090] 实验中使用的系统(52)图解显示于图3，其具有上文描述的相同的基本部分，并大致上由Meinhard雾化器(54)，收集大飞沫的雾化室(56)，渐缩喷嘴(58)组成，其中Meinhard雾化器具有混合液体细菌悬浮液进料和分散气体(N2)的同轴的喷嘴，渐缩喷嘴具有出口直径d = 0.5mm,气流通过渐缩喷嘴垂直于收集盘(60)。 [0090] The system used in this experiment (52) shown in FIG. 3 illustrates, having the same basic parts described above, and by a substantially Meinhard nebulizer (54), collecting large droplets of the spray chamber (56 ), the tapered nozzle (58), wherein a mixed liquid having a Meinhard nebulizer bacterial suspension and dispersing feed gas (N2) coaxial nozzle, the nozzle having a tapering outlet diameter d = 0.5mm, air flow through the tapering nozzle perpendicular to the catch tray (60).收集盘自身由平表面(62)组成，平表面(62)具有一个0.5_引向收集贮器(66)的孔(64)。 Collecting tray itself by a flat surface (62), with a flat surface (62) having a 0.5_ aperture (64) towards the collection reservoir (66).虽然不必须，在这个实施方案中，撞击器还包括间隔装置(68)和支持物(70)，其允许盘(60)至喷嘴(58)的距离(x)的变化从O变化到2mm(见图3插图)。 Although not necessarily, in this embodiment, the striker further comprising spacer means (68) and the support (70), which allows the plate (60) changes to a nozzle (58) a distance (x) is changed from O to 2mm ( Figure 3 inset).如之前，收集DNA于缓冲液中，其注入可能是分离的或整合的传感器系统的裂解后步骤。 As before, the DNA is collected in a buffer, which may be implanted after lysis isolated or integrated sensor system step.

[0091] 最后，本发明的系统的系统动力学表达为如下: [0091] Finally, the system according to the present invention, kinetic system is expressed as follows:

[0092] [0092]

[0093] 其中f是保持完整的细胞的比例，η d是空气中的DNA的收集效率。 [0093] where f is the proportion of cells remain intact, η d is the collection efficiency of the air DNA.在气相中，死容积大约为O。 In the gas phase, a dead volume of about O.因此，破碎是一个稳定的状态过程，其减少本发明的系统的反应时间。 Thus, the crushing process is a steady state, which reduces the reaction time of the system according to the present invention.这增强了系统的理论反应。 This increases the theoretical response of the system.EQ1&2(等式1&2)是以z' = Az+Bw的形式,其中w和z分别是时间依赖的输入和输出。 EQ1 is & 2 (Eq. 1 & 2) is z '= Az + Bw form, wherein w and z are the time-dependent input and output.

[0094] 下表2提供在EQ.1(等式I)和EQ.2(等式2)的模拟中使用的一组参数。 2 provides a set of parameters used in the simulation EQ. 1 (Equation I) and Eq. 2 (Equation 2), [0094] in the following table.对于图2a的系统，提取器是可以在30秒内裂解50 %的细胞的微射流声波仪。 For the system of FIG. 2a, the extractor can be cleaved 50% of the cells in a microfluidizer sonicator in 30 seconds.(见TCMarentis等人，Ultrasound in Medicine and Biology, 31 (9):1265_1277，(2005)，这个公开内容在此以引用并入)。 (See TCMarentis et al., Ultrasound in Medicine and Biology, 31 (9): 1265_1277, (2005), the disclosure of which is incorporated herein by reference).V1和V2分别是图2a中的第一部件和第二部件的死容积。 V1 and V2 are the dead volume FIG 2a first and second member.要理解，对于本发明的系统(图2b)，只存在一个死容积。 It is to be understood, for a system (FIG. 2b) of the present invention, there is only a dead volume.(见W.Martens-Habbena和H.Sass, AppliedandEnvironmental Microbiology, 72 (I):87_95，(2006)，这个公开内容在此以引用并入。) (See W.Martens-Habbena and H.Sass, AppliedandEnvironmental Microbiology, 72 (I): 87_95, (2006), the disclosure of which is incorporated herein by reference.)

[0095] [0095]

[0096] [0096]

[0097] 使用MATLAB®代码评估已有系统(图2a，EQ.1(等式1))，和本发明的系统(图2b，EQ.2(等式2))的时间反应(yi(t))。 [0097] The code evaluator using MATLAB® existing system (FIG. 2a, EQ.1 (Equation 1)), and a system (FIG. 2b, EQ.2 (Equation 2)) of the present invention, the reaction time (yi (t )).为了理论地评价本发明的系统提供的改进的幅度，选择产生最佳情况的操作的已有系统的参数数值(细节如上文表2)，而不是报道的对于单一已有系统的参数数组或跨越许多已有系统的平均参数数值。 Parameter values ​​(detailed above in Table 2), rather than the reported existing systems In order to evaluate the magnitude of the improved system of the present invention to provide theoretical, the selection operation to produce the best conditions for a single parameter array or across existing system many average parameter values ​​of existing systems.(见例如上文引用的TCMarentis等人&W.Martens-Habbena和H.Sass)。 (See e.g. et al cited above TCMarentis & W.Martens-Habbena and H.Sass).图4提供数据图，显不提供一步输入(a)和一个脉冲输入(b)的情况下，传统系统和本发明的系统的理论反应。 FIG 4 provides data, which was not the case to provide input step (a) and a pulse input (b), the theoretical reaction of the conventional system and the system of the present invention.(实线表示有参数的传统系统，而虚线显示本发明的系统的反应曲线，其描绘了3个61^的情况。) (Solid line represents the traditional system parameters, and the dashed curve shows the reaction system of the present invention, depicting the case where 3 ^ 61.)

[0098] 在所有的3个情况中，参数11^和V1是固定的。 [0098] In all three cases, the parameter ^ and V1 11 is fixed.f的数值从之前的工作中获得，而L和V1的数值设置在已有系统的说明书中。 F values ​​obtained from previous work, the values ​​of L and V1 provided on the existing system specification.G和η&的第一种情况相应于表2中所示的斜体字。 G and η & case corresponding to Table 2 shown in italics.如图4a所示，本发明的系统在G Jid = 292条件下操作具有达到95 %的稳定状态数值的反应，其比传统系统快多于12min。 As shown in FIG 4a, the operating system of the present invention having the reaction reaches a steady state value of 95% at a G Jid = 292 conditions, the system is faster than the traditional than 12min.而且，图4a和4b显示在I分钟标记处的信号G1)的增强，其超过29倍。 Further, FIGS. 4a and 4b show enhanced signal at the G1 I minute mark), which is more than 29 times.在第二种情况下，G Iid被设置于266以将已有系统的稳定状态的信号与步骤输入匹配。 In the second case, G Iid is provided in step 266 to signal the existing system steady state input matching.本发明的系统具有一个比已有系统更快的反应时间(7分钟15秒)，并在第一分钟内扩增信号约27倍。 The system of the present invention has a faster response time than the existing system (7 minutes and 15 seconds), and approximately 27-fold amplification of the signal within the first minute.第三种情况下，具有G Iid= 180，代表具有稳定状态数值的系统，其落后于传统系统。 In the third case, with G Iid = 180, the system represents a steady state value, which is behind the traditional system.然而，即使具有缺陷的G ηd，本发明的系统在第一分钟内仍然优于已有系统18倍。 However, even if having defects G ηd, the system according to the present invention in the first minute still 18 times better than the existing system.

[0099] 如所示，已有系统在输入信号之后约5分钟时产生最大信号，与使用现有应用的系统进行完全检测循环的15分钟的实验报道值一致。 [0099], the prior system generates the input signal, such as after about 5 minutes maximum signal, consistent with the complete detection cycle system using existing applications 15 minute experimental values ​​reported.因此，如果在一些应用中需要达到低于I分钟的检测，时间节省是重要的。 So, if you need to reach in some applications less than I minute detection, time-saving is important.(见J.Vitko, Technical Report, The NationalAcademies Press，(2005)，这个公开内容在此以引用并入)。 (See J.Vitko, Technical Report, The NationalAcademies Press, (2005), the disclosure of which is incorporated herein by reference).而且，信号的增强允许现有的传感器在检测限(LOD)内更大的灵活性。 Further, enhanced signal allows more existing sensors in the limit of detection (LOD) flexibility.

[0100] 实施例2:细胞破裂的研究 2 [0100] Example: Study of cell rupture

[0101] 使用在实施例1中描述的实验性仪器进行了研究，以显示，经过空气动力冲击的空气传播的细菌通过经历由于气体速度的突然变化引起的相对的减速而破裂。 [0101] Using the experimental apparatus described in Example 1 were studied to show, through airborne bacteria aerodynamic impact broken due to a relatively sudden change in deceleration caused by the gas velocity through experience.当X = P1/P0 < 0.53时(P1是喷嘴下游的压力，Ptl是喷嘴上游的压力)，通过以音速操作撞击器喷嘴产生空气动力的冲击。 When X = P1 / P0 <0.53 when (P1 is the pressure downstream of the nozzle, Ptl is the pressure upstream of the nozzle), the impact generated by the aerodynamic operation of striker sonic nozzle.在垂直于加速方向的细菌的表面上以波的形式的不受控制的不稳定性导致细菌细胞的破裂。 Bacteria on the surface perpendicular to the direction of acceleration to uncontrolled instabilities of the wave form of lead to rupture of the bacterial cell.临界加速ac由下列等式给出: The critical acceleration ac is given by the following equation:

[0103] 其中σ是细菌的表面张力，\是细菌的直径，05是细菌的密度。 [0103] where σ is the surface tension of bacteria, \ is the diameter of the bacteria, the bacteria density is 05.(见例如DDJoseph 等人，International Journal of Multiphase Flow,25(6-7): 1263-1303, (See e.g. DDJoseph et al., International Journal of Multiphase Flow, 25 (6-7): 1263-1303,

(1999) ;Sislian(2009)&Sislian(2010)，这些公开内容的每一个均在此以引用并入。 (1999); Sislian (2009) & Sislian (2010), the disclosures of each of which is incorporated herein by reference.) )

[0104] 对于两个不同的测试细菌气溶胶的临界加速显示于下表3，其显示大肠杆菌和萎缩芽孢杆菌(B.atoopheus)的细胞性质和诱导细菌破裂所需的临界冲击的性质。 [0104] for two different tests the bacterial aerosol Table 3 show the critical acceleration at which the display properties of E. coli cells and Bacillus atrophy (B.atoopheus) and induction of critical properties required to break bacterial shock.报道 Report

[0108] 实施例1提供的实验的设备还可以被用于测量系统的收集效率(nd)。 [0108] Experiment 1 apparatus provided in Example can also be used for collecting efficiency measuring system (nd).在如此的一个实验中，可能使用毛细管雾化器(TR-30-A1，Meinhard Glass产品)以氮气流速0.2mL/min使细菌的悬浮液成气溶胶。 In such a test, it may be using a capillary nebulizer (TR-30-A1, Meinhard Glass Products) at a nitrogen flow rate 0.2mL / min so that the bacterial suspension to the aerosol.悬浮液浓度和流速将被控制以在气溶胶中生产单一的细菌细胞。 Suspension concentration and flow rate will be controlled to produce a single bacterial cells in the aerosol.

[0109] 将使用大肠杆菌作为测试气溶胶，因为(I)大肠杆菌不要求生物安全室和(2)大肠杆菌易于培养和清洗。 [0109] E. coli is used as the test aerosol, because (I) in claim biosafety chamber E. coli and (2) culturing E. coli and easy cleaning.此外，肺炎链球菌是植物生长的细菌，预期具有与大肠杆菌vs.萎缩芽孢杆菌孢子类似的ac，并且因此具有类似的f值。 In addition, S. pneumoniae is a bacterium plant growth, E. coli vs. expected to have similar shrinkage bacillus spores ac, f and therefore have similar values.将使用PicoGreen荧光染料(P11495，LifeTechnologies)使用生产商和其他工作中提供的程序对我们的样品中的ds-DNA(双链)染色。 The use PicoGreen fluorescent dye (P11495, LifeTechnologies) using the program producers and other work provided (double-stranded) staining ds-DNA in our sample.(见上文引用的W.Martens-Habbena 和H.Sass。) (See reference above W.Martens-Habbena and H.Sass.)

[0110] 在激发波长为480nm和发射波长为520nm下使用分光荧光仪(Q32857，LifeTechnologies)可以获得ds_DNA的质量浓度形式的定量数据(低至25pg/mL)。 [0110] at an excitation wavelength of 480nm and an emission wavelength fluorescence spectroscopic analyzer (Q32857, LifeTechnologies) concentration quantitative data can be obtained in the form of mass ds_DNA (down to 25pg / mL) was used at 520nm.已知浓度的纯化的大肠杆菌ds-DNA溶液在不同稀释下被染色以获得发射强度相对于质量浓度的标准曲线。 Known concentrations of purified E. coli ds-DNA solution was dyed at different dilutions to obtain the emission intensity relative to a standard curve of concentration.将测量设备中来自于不同点的下列样品的ds-DNA(见图3:(1)注入雾化器的细菌悬浮液(PG1), (2)与⑴相同但声波处理释放内部的dS-DNA(PG2)，(3)雾化后的样品(PG3),⑷撞击器操作后的样品(PG4), (5)与(4)相同但经声波处理的(PG5)和(6)缓冲液作为空白)。 The following samples from different points of measurement apparatus ds-DNA (see FIG. 3: (1) injection nebulizer bacterial suspension (PG1), (2) the same but ⑴ acoustic dS-DNA processing inside release (PG2), (3) sample (PG3) atomized, after the striking operation ⑷ sample (PG4), (5) (4) but the same sonicated (PG5) and (6) as a buffer blank).将使用0.2 μ m的聚碳酸脂滤膜过滤所有样品以俘获完整细胞并且滤液将染色。 The use of 0.2 μ m polycarbonate membrane filter to capture all of the samples stained intact cells and the filtrate.对于三种不同的操作条件X =0.98，0.50，和0.11可以如下计算1^值: For three different operating conditions of X = 0.98,0.50, 0.11 and 1 ^ values ​​can be calculated:

[0112] 其中分子是全部提取的ds-DNA，分母是冲击前的ds-DNA的总数；f是在撞击器之前系统中颗粒损失的比例，其等于0.088 Δ} 0.029； f是实验设备的性质而非撞击器的性质；f是在实施例1中描述的。 [0112] wherein the molecule is to extract all the ds-DNA, the denominator is the total number of ds-DNA prior to impact; F is the ratio of the system loss of particles prior to the striker, which is equal to 0.088 Δ} 0.029; f is the nature of the experimental apparatus rather than the nature of the striker; F in Example 1 is described.

[0113] 在这个实验中，测量PG3作为对照以评价雾化效果；测量PG1以确定在开始的悬浮液中的细胞外ds-DNA ( = PG2-PG1)。 [0113] In this experiment, as a control measure to evaluate PG3 atomization; PG1 measurements to determine the extracellular ds-DNA in the starting suspension (= PG2-PG1).将清洗细菌悬浮液以减少细胞外的ds-DNA的量(PG2的<1%)以减少计算1^的误差，Π d是仅来自于破碎的细胞的ds-DNA的收集效率。 The bacterial suspension was washed to reduce the amount of ds-DNA extracellular (PG2 <1%) in order to reduce the calculation error of 1 ^, Π d is disrupted cells from only the collection efficiency of the ds-DNA.测量PG5以保证细胞外的ds-DNA高于开始的细胞外的ds-DNA。 PG5 to ensure measuring extracellular ds-DNA ds-DNA than extracellular started.

[0114] 之前已使用计算机计算了细菌颗粒的收集效率(η)。 It has been calculated using a computer bacterial particle collection efficiency ([eta]) prior to [0114].(见上文引用的Sislian，(2010))。 (See reference above Sislian, (2010)).使用具有标准k-瑞流模型的稳定状态Navier Stokes等式以解决气体动力学。 Using steady state Navier Stokes equations Swiss with standard k- flow model to address aerodynamic.颗粒动力学等式在低气溶胶浓度时与气体动力学等式单向偶联。 Particle dynamics equations at low aerosol concentrations and gas dynamics equations unidirectional coupling.使用随机颗粒轨迹以解释撞击器中的湍流。 Using a random particle trajectories in order to explain the turbulence striker.尽管H因子没有纳入EQ.2 (等式2)，但η是几何学和撞击器操作条件的函数，如ndo区别在于颗粒的大小:细菌是I μ m而DNA颗粒是10nm。 Although not included in Eq. 2 H factor (Equation 2), η is a function of the operating conditions but the geometry of the striker, such that the difference between the size of the granules ndo: Bacterial DNA is I μ m and the particles are 10nm.

[0116] 可以使用成熟的FLUENT代码计算收集效率(η d)。 [0116] can be calculated collection efficiency (η d) using established FLUENT code.在这个模拟中，在冲击而撞击器入口之后将释放10000个DNA颗粒并当作它们进入收集管的开口(图3)。 In this simulation, the striker after the impact will release the inlet and 10,000 DNA particles as they enter the collection tube open (FIG. 3).细菌收集效率(n)的初步的模拟结果表明，可以在不影响amax的情况下扩展收集管的入口直径(de)以增加n。 Bacterial collection efficiency (n) of the preliminary simulation results show that the inlet diameter can be extended collection tube (de) without influence amax to increase n.当直径从0.5mm变化到Imm时，分别地，对I μ m的颗粒的收集效率从η = 0.37变化到Π = 1，而amax从5.0X IO9变化到4.4X 109m/s2。 When the diameter from 0.5mm to Imm, changes, respectively, a collection efficiency of particles I μ m from η = 0.37 is changed to Π = 1, and amax is changed to from 5.0X IO9 4.4X 109m / s2.几何学的变化不影响细菌颗粒通过冲击的减速；因此将不影响f的值(古f(de))。 Geometry variations do not affect particle reduction by bacterial shock; thus will not affect the value of f (Old f (de)).因此，可以模拟多个nd= nd[x/d,de, X]以获得单喷嘴几何学，其使具有最优的G IId = 266L/min的撞击器的设计成为可能。 Accordingly, a plurality of analog nd = nd [x / d, de, X] to obtain a single nozzle geometry, which makes having optimum G IId = 266L / min of the striker designs.

[0117] 为了达到Gnd = 266L/min，变化的撞击器的阈值GnjP nd必须足够大。 [0117] In order to achieve Gnd = 266L / min, the change in threshold striker GnjP nd must be large enough.事实上，G必须大于Gnd = 266L/min因为η d < I。 Indeed, G must be greater than Gnd = 266L / min because η d <I.在一个实施方案中，可以通过并行操作多个单喷嘴几何学改进G例如如在上文实施例1中描述的那样。 In one embodiment, for example, G can be improved geometry as described above in Example 1 as described in the plurality of operations in parallel by a single nozzle.为了使喷嘴并行，气流结构不应该相互作用，因此使每一个喷嘴作用如其没有相邻的喷嘴。 Parallel to the nozzle, the air flow structure should not interact, so that each nozzle as it does not effect adjacent nozzles.这保证保持恒定。 This is guaranteed to remain constant.例如，当喷嘴直径d = Imm (每个喷嘴9.4L/min)时,需要36个喷嘴以达到G = 325L/min。 For example, when the nozzle diameter d = Imm (each nozzle 9.4L / min), the nozzle 36 to achieve the required G = 325L / min.如果计算的分离是3mm(将由两个喷嘴的模拟确定)被安排在一个正方形内，贝U需要2.1cmX 2.1cm的区域。 If the calculated separation is 3mm (determined by the analog two nozzles) are arranged in a square, the region required 2.1cmX 2.1cm shell of U.这不改变撞击器的全部直径。 All this does not change the diameter of the impactor.仅有的改变将在于泵能力的需求，可以将下游压力乘以流速来容易地估计泵能力的需求。 The only change is that the pump capacity demand, the downstream pressure may be multiplied by the flow rate of the pump to be easily estimated capacity requirements.

[0118] 实施例4:理论的TB研究 Theoretical TB: the 4 cases of [0118] Embodiment

[0119] 下面的讨论将使用肺结核(TB)作为实施例以描述本发明的气溶胶生物收集器/分析器系统的优势。 [0119] The following discussion will use the tuberculosis (TB) as the description of the present invention to biological aerosol collector / analyzer system advantages embodiment.然而，要理解，描述的基本的概念对其他细菌疾病是同等可应用的。 However, to understand the basic concepts described for other bacterial diseases are equally applicable.TB属于全世界疾病和死亡的主导病因，其在2008年导致大约130万的死亡。 TB disease belongs to the world and the leading cause of death, which causes about 1.3 million deaths in 2008.根据世界卫生组织(WH0)，现在三分之一的世界人口被TB感染并且每秒中某人将新被感染。 According to World Health Organization (WH0), now a third of the world's population is infected with TB every second and someone new is infected.在发展中国家中，痰涂片显微术(SSM)是最普遍的诊断方法。 In developing countries, sputum smear microscopy (SSM) is the most common method of diagnosis.SSM通常具有0.97的特异性，0.51的敏感性，和2-7 天的反应时间(见例如Keeler, E 等人，Nature Reviews !Diagnostics, 61006:49-57，(2006)，这个公开内容在此以引用并入)。 SSM is usually specific to 0.97, sensitivity 0.51, and 2-7 days of reaction time (see for example Keeler, E, et al., Nature Reviews Diagnostics, 61006:! 49-57, (2006), the disclosure of which is incorporated by reference).慢反应时间导致患者跟踪的损失。 Slow response time resulting in loss of patient tracking.因为痰的收集是个多步骤的过程，一些成人样品被唾液稀释并包含不足量的结核分枝杆菌细菌。 Because the collection of sputum is a multi-step process, some adult saliva samples were diluted and contains an insufficient amount of Mycobacterium tuberculosis bacteria.因此可以提供更好的特异性，敏感性，反应时间和样品收集的方法将极大加强一个可治愈疾病的诊断。 Therefore can provide better specificity and sensitivity of the method, the reaction time and sample collection will greatly strengthen a diagnosis of incurable disease.

[0120] 此外，从占TB阳性人群的11%的儿童中获得痰是困难的，并要求特殊的步骤例如咳痰、胃抽吸，和痰诱导；所有这些对于健康护理的提供者和儿童都是不愉快。 [0120] In addition, sputum obtained from accounting for 11% TB-positive population of children is difficult and requires special steps such as sputum, gastric aspiration and sputum induction; for all these health care providers and children alike It is unpleasant.而且，无论何时可获得的，这些样品的质量也将低于从成人获得的样品质量。 Moreover, whenever available, the quality of these samples will also be less than the mass of the sample obtained from adults.通过消除痰收集的多步骤过程，本发明的空气传播的细菌检测系统还减少从成年患者中收集的样品质量的变化。 Multi-step process by eliminating phlegm collected airborne bacteria detection system of the present invention also reduces the change in mass of the sample collected from adult patients.而且，在不发达区域的患者常常没有回到诊所跟踪的方法(所有患者中的10-15% )。 Moreover, patients in developed regions often do not come back to the clinic to track (10-15% of all patients).通过结合样品的收集和结核分枝杆菌DNA的检测，本发明的设备将可以在大约几分钟到几小时间获得TB读数，这消除跟踪的损失，导致大约95000个年调整生命拯救(Annual AdjustedLives Saved (AALS))(再次见上文引用的Keeler (2006))。 By collecting and detecting binding of the DNA samples of Mycobacterium tuberculosis, the apparatus of the present invention will be between about a few minutes to several hours to obtain readings TB, which eliminates the loss of tracking caused approximately 95,000 Adjustment lifesaving (Annual AdjustedLives Saved (AALS)) (see the above cited again Keeler (2006)).此外，生物传感器将还提供相比于标准的SSM技术更好的敏感性和特异性。 Further, the biosensor further provides SSM compared to standard techniques better sensitivity and specificity.使用新设备的AALS的全部潜力为625000，是超过现在的方法的显著的改进.[0121] 再次使用TB作为实施例，一个典型的TB患者随每一次咳嗽或打喷嚏分别产生大约3000和40000个包括结核分枝杆菌的感染性的飞沫。 Full potential AALS using the new equipment to 625,000, more than the present method is significantly improved. [0121] Again using TB as an example, a typical TB patient with every cough or sneeze respectively generated approximately 3000 and 40,000 including infectious droplets Mycobacterium tuberculosis.如图1al和la2中描绘的，这些飞沫(16)作为开始的样品，用于使用本发明的气溶胶生物收集器/分析器系统来诊断TB患者。 FIG 1al and la2 depicted, these droplets (16) begins as a sample, the present invention is used for biological aerosol collector / analyzer system to diagnose TB patients.根据这些统计，四个咳嗽的平均可以产生足够的细胞收集于Iml以超过现在的SSM的检测限(> 10000细胞/ml)。 According to these statistics, an average of four cough may produce sufficient cells were collected in Iml SSM exceeding current limits of detection (> 000 cells / ml).确定现在的系统对于一个测试情况如TB的效率的一个方法可包括如实施例1描述的具有4个雾化器的仪器(以模拟4次咳嗽)上进行实验，产生大约12000个飞沫。 A system for determining the current situation as a method of testing the efficiency of the TB may include instruments as described in Example 1 of the embodiment having four atomizer (to simulate coughs 4) on the experiment, produces approximately 12,000 droplets.在如此的一个实验中，将测量ds-DNA的质量浓度(y)并记录每一次运行。 In such a test, the measurement of the concentration of ds-DNA of (y) and recorded every run.对于注入雾化器的悬浮液中细菌的每一个浓度(Xi，I < i < c)，将执行多个运行(N)。 For injection nebulizer suspension each concentration (Xi, I <i <c) bacteria, a plurality of execution run (N).可能变化浓度(xi)以模拟有TB和没有TB的患者。 Possible changes in the concentration (xi) to simulate patients with TB and TB are not.可能基于衍生的整个的实验数据设置的特异性和敏感性选择ds-DNA浓度的阈值，以决定给出一个与输入浓度匹配的读数的全部运行(N)的比例(N')(对于Xi = O细胞/ml，匹配的读数是低于阈值的ds-DNA的质量浓度(y))。 Specific threshold value may be based on the proportion of the entire experimental data derived and sensitivity settings selection ds-DNA concentration, with a given input to determine the concentration of all readings matching operation (N), (N ') (for Xi = O cells / ml, matching the reading is lower than the concentration of ds-DNA threshold (y)).在一个实施方案中，可以使用足以建立TB样品收集的功效的无区分的染料。 In one embodiment, the sample may be used to collect enough to establish the efficacy of TB indiscriminate dye.在一个更加先进的案例中，可能使用一个特异性的生物传感器以辨别不同细菌。 In a more advanced cases, it may use a specific biosensors to identify different bacteria.此外，可以整合收集和生物传感器模型，和使用用于人类测试的面罩以替换雾化器 In addition, the collection and the biosensor may be integrated model, and use of a mask used in humans to replace the test atomiser

[0122] 概述 [0122] Overview

[0123] 总的来说，描述了气溶胶生物收集器/分析器系统，其在每一个可测量的方面均显示超过传统系统的显著改进的表现。 [0123] In general, it describes a biological aerosol collector / analyzer system, than conventional systems which showed significant improvement in the performance of each of a measurable aspect.很好的被接受，通过如下评价如此的生物传感器系统的表现:(I)提取和传递IC至传感器的时间和效率，(2)干扰裂解后步骤，(3)花费和(4)直接与患者对接的能力。 Is well accepted by the evaluation of such a biosensor system performance: (I) to the IC extraction and delivery time and efficiency of the sensor, (2) interfere with the cleavage step, (3) cost, and (4) directly with the patient docking capability.在以上实施例1到3提供的讨论中处理了第一点。 Processing the first point in the discussion of Examples 1 to 3 provided in the above embodiment.特别的，本发明的设备直接收集至撞击器，没有其他室存在于患者和撞击器之间。 In particular, the apparatus of the present invention is collected directly to the striker, no additional chamber is present between the patient and the striker.这允许细菌的足够浓度被收集。 This allows a sufficient concentration of bacteria is collected.

[0124] •本发明的设备不通过大小分开空气颗粒。 Equipment [0124] • the present invention is not size particles are separated by air.这允许要被收集的颗粒在一个阶段而不是分开相同的样品于不同的阶段，提供在相同的患者的流出量下更浓缩的样品。 This allows particles to be collected in a separate phase instead of the same sample at different stages, a more concentrated at the outflow of the same patient sample.

[0125] •本发明的设备直接收集细菌和细菌的组分于液体样品中。 Equipment [0125] • the present invention is collected directly bacteria and bacterial components in the liquid sample.这允许样品的轻松提取和避免样品暴露导致的污染。 This allows for easy extraction of the sample and to avoid sample contamination caused by exposure.这还允许批取样和在线取样。 This also allows batch sampling and online sampling.

[0127] •与传统的两室系统相比，本发明的系统是紧凑和简单的。 [0127] • Compared with the conventional two-chamber system, the system of the present invention is compact and simple.

[0128] •最后，传统的系统通常要求使用化学的方法，其要求试剂的贮藏和在一些情况中使用专门的微射流。 [0128] • Finally, conventional systems typically require the use of a chemical method, which requires the use of special storage agent and microjets in some cases.使用微射流的小型化减少死容积(O但其增加花费。此外，一些微射流系统产生局部加热，其可使蛋白质变性。而且，需要以批模式操作它们，这可以在任何地方花费30秒到4分钟。 Using microfluidics reduce the size of the dead volume (O but its cost increases. Moreover, some microfluidic system generating localized heating, which can denature the protein. Further, they are required to operate in batch mode, it can take 30 seconds to anywhere 4 minutes.

[0129] 等同声明 [0129] equivalent statement

[0130] 已出于阐述和描述的目的，展示了本发明的这一描述。 [0130] and has been described for purposes of illustration, showing the description of the present invention.不旨在彻底详尽，也不旨在限制本发明于描述的精确的形式，并且由于上文的教导，很多修改和变化是可能的。 It is not intended to completely exhaustive, nor is it intended to limit the invention to the precise form described, and since the above teachings, many modifications and variations are possible.选择和描述实施方案以最好地解释本发明的原理及其实践的应用。 Embodiments were chosen and described to best explain the principles and practice of the present invention is applied.这个说明书将使其他本领域技术人员能够以各种实施方案最好地利用和实践本发明，并作出各种合适于特殊使用的修改。 This description will enable others skilled in the art to best utilize the various embodiments and practice the invention, and that various modifications appropriate to the particular use.由下面的权利要求限定本发明的范围。 Defined by the scope of the invention claimed in the following claims.

Claims (26)

Translated from Chinese

1.一种气溶胶生物收集器/分析器系统，包括: 患者接口，其具有第一终端和第二终端并限定其间的流体通道，其中所述第一终端能够接合患者以至于所述接口捕获患者的呼气流出物的至少一部分作为气体样品，所述患者的呼气流出物的至少一部分包括其中包含的任何生物微粒； 相互连接管，其具有第一终端和第二终端并限定其间的流体通道，其中所述相互连接管的第一终端接合所述患者接口的第二终端并与所述患者接口的第二终端互锁； 空气动力撞击器，其具有第一终端和第二终端并限定其间的流体通道，其中所述撞击器的第一终端接合所述相互连接管的第二终端并与所述相互连接管的第二终端互锁，并且其中所述空气动力撞击器向所述气体样品施加惯性减速力，并且其中惯性力的强度能够变化，以至于在低惯性力下，所述样品中的任何生物微粒 A biological aerosol collector / analyzer system, comprising: a patient interface having a first end and a second end and defining a fluid passage therebetween, wherein the first terminal is able to engage a patient such that said interface captures at least a part of the gas sample, at least a portion of the patient's exhalation breath effluent effluent patients include any biological particles contained therein; interconnecting tube having a first end and a second end and defining therebetween a fluid channel, wherein a first terminal connected to said another terminal of said second tube engaging patient interface and the patient interface with a second terminal of the interlock; aerodynamic striker, having first and second terminals and defining a fluid passage therebetween, wherein a first terminal of said striker engages said second terminal are interconnected to each other with the tube and a second end interlocking connection tube, and wherein said aerodynamic striker to the gas coasts sample application force, inertial force and the intensity of which can vary, so that at low inertial force, the sample is any biological particle完整地通过所述撞击器，并且在高于阈值的惯性力下，所述样品中的任何生物微粒被裂解以释放其内部组分； 收集容器，其具有第一终端和第二终端，且限定容纳流体的主体，其中所述收集容器的第一终端接合所述撞击器的第二终端并与所述撞击器的第二终端互锁，其中液体介质被置于容纳流体的主体内，所述液体介质用于保存来自于所述气体样品的完整的细菌微粒或生物微粒的内部组分两者之一，并且其中形成了与所述容纳流体的主体流体连通的出口，以允许将材料收集在所述容纳流体的主体中；和泵，其与所述系统流体连通，用以产生从所述接口的第一终端到所述收集容器的流压，以至于所述气体样品被推动通过由所述相互连接管和所述撞击器限定的所述流体通道而进入所述收集容器。 To release its entirety by the internal components of the striker, and the inertial force above a threshold value, any biological particles in the sample are lysed; collection container, having a first terminal and a second terminal, and defining containing body fluid, wherein the first terminal of the second terminal collection container engages said striker and interlock with the second terminal of the striker, wherein the liquid medium is disposed within the receiving body fluid, said a liquid medium for storing one of both of the inner component intact bacterial particles or biological particle from the gaseous sample, and an outlet formed therein in communication with said fluid containing body fluid to allow the material collected in the the receiving fluid in the body; and a pump, in fluid communication with said system for generating a flow pressure to the collection container from a first terminal of the interface, so that the gas sample is being pushed by the interconnecting said pipe and said striker defining the fluid passage into the collection container.

2.根据权利要求1所述的系统，其中所述接口包括全脸持续正气道压力面罩，所述全脸持续正气道压力面罩收集患者的全部呼出物。 2. The system according to claim 1, wherein said interface comprises a continuous positive airway pressure of a full face mask, a full face continuously whole exhalation positive airway pressure mask was collected patient.

3.根据权利要求1所述的系统，其中所述相互连接管还包括至少一个辅助补偿入口，所述至少一个辅助补偿入口被横切于所述相互连接管的轴布置，具有与所述相互连接管流体地相互连接的第一终端，和提供至大气的出口的第二终端。 3. The system of claim 1, wherein said interconnection further comprises at least one auxiliary tube inlet compensating, at least one auxiliary compensation inlet is disposed transversely to the axis of said interconnecting pipe, each having the first terminal connecting tube fluidly connected to each other, and the second terminal is provided to the outlet to the atmosphere.

4.根据权利要求3所述的系统，其中所述至少一个辅助补偿入口被布置为与所述相互连接管成大于90度的角。 4. The system of claim 3, wherein said at least one auxiliary inlet is arranged to compensate for the angle of each connecting tube is greater than 90 degrees.

5.根据权利要求3所述的系统，还包括过滤器，所述过滤器被布置于所述辅助补偿入口的第二终端出口处。 5. The system according to claim 3, further comprising a filter, the filter is disposed at the outlet of the second terminal of the auxiliary compensation inlet.

6.根据权利要求5所述的系统，其中所述过滤器是HEPA过滤器。 6. The system according to claim 5, wherein said filter is a HEPA filter.

7.根据权利要求1所述的系统，其中所述撞击器包括渐缩喷嘴和平收集表面，所述渐缩喷嘴限定所述撞击器的所述流体通道，所述平收集表面被布置于所述渐缩喷嘴的远端并垂直于所述流动通道，所述平收集表面具有布置于其中并与所述渐缩喷嘴的所述流体通道对准的入口孔，并且其中所述入口孔与所述撞击器的第二终端流体连通。 7. The system according to claim 1, wherein said striker comprises a tapered nozzle peace collection surface, the tapered nozzle of the fluid passage defining said striker, said flat being disposed on said collection surface the tapered distal end of the nozzle and perpendicular to said flow channel, said flat surface having a collection aperture disposed therein and an inlet of the tapered nozzle of the fluid passage aligned, and wherein said inlet aperture and said a second end in fluid communication with the striker.

9.根据权利要求7所述的系统，其中所述喷嘴到所述收集表面的距离(X)和所述喷嘴的直径⑷具有下述比x/d = 1.2。 9. The system of claim 7, wherein said nozzle distance (X) of the collection surface and the nozzle has a diameter ⑷ ratio x / d = 1.2.

10.根据权利要求7所述的系统，其中所述喷嘴的面对样品的表面是镜面抛光的。 Facing surface of the sample 10. The system of claim 7, wherein said nozzle is mirror-polished.

11.根据权利要求7所述的系统，其中所述撞击器包括多于一个喷嘴。 11. The system of claim 7, wherein said striker comprises more than one nozzle.

12.根据权利要求1所述的系统，其中所述收集容器的第二终端向所述出口逐渐减小。 12. The system according to claim 1, wherein the second terminal of the collection container is gradually reduced toward the outlet.

13.根据权利要求1所述的系统，其中所述收集容器的第一终端和所述液体介质的表面之间的距离是约4_。 13. The system according to claim 1, wherein a distance between the first terminal and the surface of the liquid medium of the collection container is approximately 4_.

14.根据权利要求1所述的系统,其中所述出口允许对所述容纳流体的主体中的材料取样，而不打开所述系统的任何其他部分。 14. The system according to claim 1, wherein said outlet for allowing the receiving body fluid sample of a material, without opening any other part of the system.

15.根据权利要求14所述的系统，其中通过注射器进入所述出口。 15. The system according to claim 14, wherein the syringe enters through the outlet.

16.根据权利要求1所述的系统，其中所述液体介质是缓冲液。 16. The system according to claim 1, wherein said liquid medium is a buffer.

17.根据权利要求1所述的系统，还包括能够检测样品中的细菌微粒的传感器，所述传感器被布置成与所述收集容器流体连接。 17. The system according to claim 1, further comprising a sensor capable of detecting bacteria in a sample of microparticles, the sensor is arranged in connection with the fluid collection container.

18.根据权利要求1所述的系统,其中所述样品中的细胞的内部组分包括DNA。 18. The system according to claim 1, wherein the internal components of the cell sample comprises DNA.

19.根据权利要求1所述的系统，其中所述系统的组件间的相互连接部包括具有阀门的快速分离偶联器，以至于一旦分离，所述阀门自动关闭。 19. The system according to claim 1, wherein the interconnecting portion between the components of the system comprises a valve having a quick disconnect coupling device, such that once separated, the valve automatically closes.

20.根据权利要求1所述的系统，其中所述系统的组件是可灭菌的。 20. The system according to claim 1, wherein the components of the system is sterilized.

21.根据权利要求1所述的系统，其中所述系统的面对样品的表面由塑料材料形成。 21. The system according to claim 1, wherein the surface of the sample facing the system formed from a plastic material.

22.根据权利要求1所述的系统，其中所述样品由选自呼吸、咳嗽和打喷嚏的方法形成。 22. The system according to claim 1, wherein said sample is selected from The method of breathing, coughing and sneezing are formed.

23.—种针对细菌感染来收集和分析气溶胶样品的方法，包括:` 收集患者的流出物作为样品，所述患者的流出物包括其中包含的任何生物微粒； 传输所述样品至撞击器； 选择待施加于所述样品的惯性力的强度，其中所述强度能够变化，以至于所述样品中的任何细菌微粒完整通过所述撞击器，或以至于所述样品中的任何细菌微粒被裂解以释放其内部组分； 向所述样品施加选择的惯性力；和将完整的生物微粒或生物微粒的内部组分两者之一收集于流体介质中，作为分析物溶液用于分析。 23.- ways to aerosol samples collected and analyzed for bacterial infection, comprising: `effluent was collected as a sample of a patient, the patient effluent comprises any biological particles contained therein; transmitting the samples to the striker; selecting an inertial force to be applied to the intensity of the sample, wherein the intensity can be varied, so that any bacteria in the sample particles complete by the striker, so that any bacteria or particles in said sample is cleaved to release its internal components; inertial force is applied to the selected sample; and one of both of the inner component or intact biological particles collected biological particles in a fluid medium, as analytical solution for analysis.

24.根据权利要求21所述的方法，还包括针对患者的震动流出物补偿所述系统的压力。 24. The method of claim 21, further comprising a pressure shock for the patient compensate for the effluent system.

25.根据权利要求21所述的方法，还包括实时分析所述分析物溶液中的细菌污染物。 25. The method of claim 21, further comprising analyzing said real-time analysis of bacterial contaminants in solution.